Hepatic glucokinase turnover in intact and adrenalectomized rats in vivo.
Regulation of synthesis and degradation of glucokinase, key enzyme of glucose metabolism in liver, was investigated in intact and adrenalectomized rats in vivo, using pulse-labeling experiments and a specific antibody against the enzyme. Refeeding glucose in starved rats resulted in a marked rise in glucokinase activity (from the starvation value 4.8 mU/mg protein to 9.6 mU/mg protein at 4 h, and to 21.8 mU/mg protein at 8 h), which closely correlated to the increase in enzyme synthesis by factor 1.7 at 4 h and 4.1 at 8 h. Similar alterations in enzyme activity and synthesis were observed after glucose refeeding in adrenalectomized/glucocorticoid-restored rats. In contrast, refeeding glucose in adrenalectomized rats led within 8 h only to a small elevation in enzyme activity (from the starvation value 4.2 mU/mg protein to 9.6 mU/mg protein) and a minor rise in enzyme synthesis (factor 1.7). Glucocorticoids per se were without effect on glucokinase activity and synthesis in starved rats. When adapted to pure glucose diet, intact, adrenalectomized and adrenalectomized/glucocorticoid-restored rats showed highly elevated levels in glucokinase (27, 23, 28 mU/mg protein, respectively). However, enzyme synthesis was elevated significantly only in intact and adrenalectomized/glucocorticoid-restored rats. Under these conditions glucokinase degradation was estimated by the double-pulse-labeling technique, applying [14C]leucine and [3H]leucine. From the 3H/14C ratios the apparent half-lives were calculated: 17 h in intact and adrenalectomized/glucocorticoid-restored rats, and 35 h in adrenalectomized rats. It is concluded that in vivo glucocorticoids not only exert a 'permissive' action on glucokinase induction, but also enhance the degradation of the enzyme.
D000315:Adrenalectomy; D000818:Animals; D000906:Antibodies / Q000276:immunology; D005941:Glucokinase / Q000096:biosynthesis / Q000276:immunology / Q000378:metabolism*; D005947:Glucose / Q000494:pharmacology; D006854:Hydrocortisone / Q000494:pharmacology; D005779:Immunodiffusion; D008099:Liver / Q000187:drug effects / Q000201:enzymology*; D008297:Male; D051381:Rats; D013217:Starvation
D000906:Antibodies; D005941:Glucokinase; D005947:Glucose; D006854:Hydrocortisone